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1.
Proc Natl Acad Sci U S A ; 120(48): e2309412120, 2023 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-37983500

RESUMO

Bunyaviruses are enveloped negative or ambisense single-stranded RNA viruses with a genome divided into several segments. The canonical view depicts each viral particle packaging one copy of each genomic segment in one polarity named the viral strand. Several opposing observations revealed nonequal ratios of the segments, uneven number of segments per virion, and even packaging of viral complementary strands. Unfortunately, these observations result from studies often addressing other questions, on distinct viral species, and not using accurate quantitative methods. Hence, what RNA segments and strands are packaged as the genome of any bunyavirus remains largely ambiguous. We addressed this issue by first investigating the virion size distribution and RNA content in populations of the tomato spotted wilt virus (TSWV) using microscopy and tomography. These revealed heterogeneity in viral particle volume and amount of RNA content, with a surprising lack of correlation between the two. Then, the ratios of all genomic segments and strands were established using RNA sequencing and qRT-PCR. Within virions, both plus and minus strands (but no mRNA) are packaged for each of the three L, M, and S segments, in reproducible nonequimolar proportions determined by those in total cell extracts. These results show that virions differ in their genomic content but together build up a highly reproducible genetic composition of the viral population. This resembles the genome formula described for multipartite viruses, with which some species of the order Bunyavirales may share some aspects of the way of life, particularly emerging properties at a supravirion scale.


Assuntos
Orthobunyavirus , Tospovirus , Orthobunyavirus/genética , RNA Viral/genética , Tospovirus/genética , Genoma Viral/genética , Vírion/genética
2.
Mol Plant Microbe Interact ; 36(11): 705-715, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37432156

RESUMO

The NLR (nucleotide-binding leucine-rich repeat) class immune receptor Sw-5b confers resistance to Tomato spotted wilt orthotospovirus (TSWV). Although Sw-5b is known to activate immunity upon recognition of the TSWV movement protein NSm, we know very little about the downstream events that lead to resistance. Here, we investigated the Sw-5b-mediated early transcriptomic changes that occur in response to mechanical and thrips-mediated inoculation of TSWV, using near-isogenic tomato lines CNPH-LAM 147 (Sw5b+/+) and Santa Clara (Sw-5b-/-). We observed earlier Sw-5b-mediated transcriptional changes in response to thrips-mediated inoculation compared with that in response to mechanical inoculation of TSWV. With thrips-mediated inoculation, differentially expressed genes (DEGs) were observed at 12, 24, and 72 h postinoculation (hpi). Whereas with mechanical inoculation, DEGs were observed only at 72 hpi. Although some DEGs were shared between the two methods of inoculation, many DEGs were specific to either thrips-mediated or mechanical inoculation of TSWV. In response to thrips-mediated inoculation, an NLR immune receptor, cysteine-rich receptor-like kinase, G-type lectin S-receptor-like kinases, the ethylene response factor 1, and the calmodulin-binding protein 60 were induced. Fatty acid desaturase 2-9, cell death genes, DCL2b, RIPK/PBL14-like, ERF017, and WRKY75 were differentially expressed in response to mechanical inoculation. Our findings reveal Sw-5b responses specific to the method of TSWV inoculation. Although TSWV is transmitted in nature primarily by the thrips, Sw-5b responses to thrips inoculation have not been previously studied. Therefore, the DEGs we have identified in response to thrips-mediated inoculation provide a new foundation for understanding the mechanistic roles of these genes in the Sw-5b-mediated resistance. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Assuntos
Vírus de RNA , Solanum lycopersicum , Tisanópteros , Tospovirus , Animais , Solanum lycopersicum/genética , Tisanópteros/genética , Tospovirus/fisiologia , Doenças das Plantas , Proteínas do Movimento Viral em Plantas/metabolismo , Vírus de RNA/metabolismo
3.
Curr Opin Insect Sci ; 56: 101021, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36925103

RESUMO

Resistant varieties are critical tools for crop production, and single-resistance genes providing strong protection against pests or pathogens are deployed in agriculture. Durability of these traits is threatened by emergence of resistance-breaking pests and pathogens. This review focuses on acylsugar-mediated resistance in tomato. Wild tomatoes have type-IV trichomes that exude chemically complex mixtures of acylsugars altering behavior and suppressing multiple pest species, and with thrips and whiteflies (WF), suppressing virus transmission, for example, Tomato spotted wilt orthotospovirus and Tomato yellow leaf curl virus, respectively. Marker-assisted selection and bioassays led to development of advanced cultivated tomato breeding lines rich in acylsugar variations, allowing acylsugar-mediated resistance to be combined with other resistance traits providing a layered defense system that reduces pest populations and virus disease prevalence. This strategy also holds promise for enhancing durability of virus resistance genes by reducing the intensity of selection for resistance-breaking variants.


Assuntos
Hemípteros , Solanum lycopersicum , Tisanópteros , Animais , Doenças das Plantas , Produção Agrícola
4.
Insect Biochem Mol Biol ; 149: 103843, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36113709

RESUMO

Successful transmission of tomato spotted wilt virus (TSWV) by Frankliniella occidentalis requires robust infection of the salivary glands (SGs) and virus delivery to plants during salivation. Feeding behavior and transmission efficiency are sexually-dimorphic traits of this thrips vector species. Proteins secreted from male and female SG tissues, and the effect of TSWV infection on the thrips SG proteome are unknown. To begin to discern thrips factors that facilitate virus infection of SGs and transmission by F. occidentalis, we used gel- and label-free quantitative and qualitative proteomics to address two hypotheses: (i) TSWV infection modifies the composition and/or abundance of SG-expressed proteins in adults; and (ii) TSWV has a differential effect on the male and female SG proteome and secreted saliva. Our study revealed a sex-biased SG proteome for F. occidentalis, and TSWV infection modulated the SG proteome in a sex-dependent manner as evident by the number, differential abundance, identities and generalized roles of the proteins. Male SGs exhibited a larger proteomic response to the virus than female SGs. Intracellular processes modulated by TSWV in males indicated perturbation of SG cytoskeletal networks and cell-cell interactions, i.e., basement membrane (BM) and extracellular matrix (ECM) proteins, and subcellular processes consistent with a metabolic slow-down under infection. Several differentially-abundant proteins in infected male SGs play critical roles in viral life cycles of other host-virus pathosystems. In females, TSWV modulated processes consistent with tissue integrity and active translational and transcriptional regulation. A core set of proteins known for their roles in plant cell-wall degradation and protein metabolism were identified in saliva of both sexes, regardless of virus infection status. Saliva proteins secreted by TSWV-infected adults indicated energy generation, consumption and protein turnover, with an enrichment of cytoskeletal/BM/ECM proteins and tricarboxylic acid cycle proteins in male and female saliva, respectively. The nonstructural TSWV protein NSs - a multifunctional viral effector protein reported to target plant defenses against TSWV and thrips - was identified in female saliva. This study represents the first description of the SG proteome and secretome of a thysanopteran and provides many candidate proteins to further unravel the complex interplay between the virus, insect vector, and plant host.


Assuntos
Tisanópteros , Tospovirus , Animais , Feminino , Flores , Masculino , Doenças das Plantas , Plantas , Proteoma/metabolismo , Proteômica , Glândulas Salivares , Tisanópteros/metabolismo , Tospovirus/fisiologia
6.
BMC Biol ; 18(1): 142, 2020 10 19.
Artigo em Inglês | MEDLINE | ID: mdl-33070780

RESUMO

BACKGROUND: The western flower thrips, Frankliniella occidentalis (Pergande), is a globally invasive pest and plant virus vector on a wide array of food, fiber, and ornamental crops. The underlying genetic mechanisms of the processes governing thrips pest and vector biology, feeding behaviors, ecology, and insecticide resistance are largely unknown. To address this gap, we present the F. occidentalis draft genome assembly and official gene set. RESULTS: We report on the first genome sequence for any member of the insect order Thysanoptera. Benchmarking Universal Single-Copy Ortholog (BUSCO) assessments of the genome assembly (size = 415.8 Mb, scaffold N50 = 948.9 kb) revealed a relatively complete and well-annotated assembly in comparison to other insect genomes. The genome is unusually GC-rich (50%) compared to other insect genomes to date. The official gene set (OGS v1.0) contains 16,859 genes, of which ~ 10% were manually verified and corrected by our consortium. We focused on manual annotation, phylogenetic, and expression evidence analyses for gene sets centered on primary themes in the life histories and activities of plant-colonizing insects. Highlights include the following: (1) divergent clades and large expansions in genes associated with environmental sensing (chemosensory receptors) and detoxification (CYP4, CYP6, and CCE enzymes) of substances encountered in agricultural environments; (2) a comprehensive set of salivary gland genes supported by enriched expression; (3) apparent absence of members of the IMD innate immune defense pathway; and (4) developmental- and sex-specific expression analyses of genes associated with progression from larvae to adulthood through neometaboly, a distinct form of maturation differing from either incomplete or complete metamorphosis in the Insecta. CONCLUSIONS: Analysis of the F. occidentalis genome offers insights into the polyphagous behavior of this insect pest that finds, colonizes, and survives on a widely diverse array of plants. The genomic resources presented here enable a more complete analysis of insect evolution and biology, providing a missing taxon for contemporary insect genomics-based analyses. Our study also offers a genomic benchmark for molecular and evolutionary investigations of other Thysanoptera species.


Assuntos
Genoma de Inseto , Características de História de Vida , Tisanópteros/fisiologia , Transcriptoma , Animais , Produtos Agrícolas , Comportamento Alimentar , Cadeia Alimentar , Imunidade Inata/genética , Percepção , Filogenia , Reprodução/genética , Tisanópteros/genética , Tisanópteros/imunologia
7.
Pathogens ; 9(8)2020 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-32764311

RESUMO

Tomato spotted wilt virus (TSWV; species Tomato spotted wilt orthotospovirus; genus Orthotospovirus; family Tospoviridae) is a thrips-transmitted virus that can cause substantial economic losses to many crops, including tomato (Solanum lycopersicum). Since 2005, TSWV emerged as an economically important virus of processing tomatoes in the Central Valley of California, in part due to increased populations of the primary thrips vector, western flower thrips (WFT; Frankliniella occidentalis). To develop an understanding of the epidemiology of TSWV in this region, population densities of WFT and incidence of TSWV were monitored in California's processing tomato transplant-producing greenhouses and associated open fields from 2007 to 2013. Thrips were monitored with yellow sticky cards and in tomato flowers, whereas TSWV incidence was assessed with indicator plants and field surveys for virus symptoms. All thrips identified from processing tomato fields were WFT, and females were three-fold more abundant on sticky cards than males. Symptoms of TSWV infection were observed in all monitored processing tomato fields. Incidences of TSWV ranged from 1 to 20%, with highest incidence found in late-planted fields. There was no single primary inoculum source, and inoculum sources for thrips/TSWV varied depending on the production region. These results allowed us to develop a model for TSWV infection of processing tomatoes in the Central Valley of California. The model predicts that low levels of primary TSWV inoculum are amplified in early-planted tomatoes and other susceptible crops leading to highest levels of infection in later-planted fields, especially those with high thrips populations. Based upon these findings, an integrated pest management (IPM) strategy for TSWV in processing tomatoes in California was devised. This IPM strategy focuses on strategic field placement (identification of high-risk situations), planting TSWV- and thrips-free transplants, planting resistant varieties, monitoring for TSWV symptoms and thrips, roguing infected plants, thrips management targeting early generations, extensive sanitation after harvest, and strategic cropping to avoid overlap with winter bridge crops.

8.
Sci Rep ; 9(1): 17157, 2019 11 20.
Artigo em Inglês | MEDLINE | ID: mdl-31748622

RESUMO

There is increasing evidence that acylsugars deter insect pests and plant virus vectors, including the western flower thrips (WFT), Frankliniella occidentalis (Pergande), vector of tomato spotted wilt virus (TSWV). Acylsugars are sugar-polyesters composed of saturated, un-saturated, and variously branched short and long chain fatty acids (FAs) esterified to a glucose (acylglucose) or sucrose (acylsucrose) moiety. We sought to understand how acylsucrose amount and composition of associated FA profiles interacted to mediate resistance to WFT oviposition and TSWV inoculation on tomato leaves. Towards this goal, we examined WFT oviposition and TSWV inoculation behavior on tomato lines bred to exude varying amounts of acylsucrose in association with diverse FA profiles. Our data show that as acylsucrose amounts increased, WFT egg-laying (oviposition) decreased and TSWV inoculation was suppressed. Western flower thrips also responded to FA profiles that included iC4, iC11, nC12 and nC10 FA. These findings support improving acylsugar-mediated resistance against WFT by breeding tomatoes exuding greater amounts of acylsucrose associated with specific FA profiles. We show that increasing acylsucrose amount output by type IV trichomes and selecting for particular FA profiles through advanced breeding profoundly affects WFT behavior in ways that benefit management of WFT as direct pests and as TSWV vectors.


Assuntos
Flores/virologia , Insetos Vetores/virologia , Oviposição/fisiologia , Folhas de Planta/virologia , Solanum lycopersicum/virologia , Tisanópteros/virologia , Tospovirus/patogenicidade , Animais , Ácidos Graxos/metabolismo , Feminino , Flores/metabolismo , Insetos/virologia , Solanum lycopersicum/metabolismo , Doenças das Plantas/virologia , Folhas de Planta/metabolismo , Sacarose/metabolismo , Tricomas/virologia
9.
Arch Virol ; 164(11): 2829-2836, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31486908

RESUMO

The complete sequence of the medium (M) and small (S) RNA genome segments were determined for twelve isolates of impatiens necrotic spot virus from eight plant species. The M- and S-RNAs of these isolates shared 97-99% and 93-98% nucleotide sequence identity, respectively, with the corresponding full-length sequences available in public databases. Phylogenetic analysis based on the M- or S-RNA sequences showed incongruence in the phylogenetic position of some isolates, suggesting intraspecies segment reassortment. The lack of phylogenetic discordance in individual and concatenated sequences of individual genes encoded by M- or S-RNAs suggests that segment reassortment rather than recombination is driving evolution of these INSV isolates.


Assuntos
RNA Viral/genética , Vírus Reordenados/genética , Tospovirus/genética , Sequência de Bases , Genoma Viral/genética , Plantas/virologia , Análise de Sequência de RNA , Tospovirus/isolamento & purificação
10.
PLoS One ; 13(7): e0201583, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30063755

RESUMO

Tomatoes (Solanum lycopersicum L.) have been bred to exude higher amounts or different types of the specialized plant metabolites, acylsugars, from type IV trichomes. Acylsugars are known to deter several herbivorous insect pests, including the western flower thrips (WFT), Frankliniella occidentalis (Pergande); however, all previous studies investigated the effect of acylsugars on leaves, or acylsugar extracts obtained from leaves. In spite of the WFT predilection for flowers, there is a gap in knowledge about flower defenses against thrips damage. This is especially important in light of their capacity to acquire and inoculate viruses in the genus Orthotospovirus, such as Tomato spotted wilt orthotospovirus (TSWV), in flowers. Therefore, we turned our attention to assessing thrips oviposition differences on flowers of 14 entries, including 8 interspecific hybrids, 5 tomato lines bred for specific acylsugar-related characteristics (type IV trichome densities, acylsugar amount, sugar moiety and fatty acid profile), and a fresh market tomato hybrid, Mt. Spring, which only produces trace amounts of acylsugars. Our results show that the density of the acylsugar droplet bearing type IV trichomes is greatest on sepals, relative to other flower structures, and accordingly, WFT avoids oviposition on sepals in favor of trichome-sparse petals. In concordance with past studies, acylsugar amount was the most important acylsugar-related characteristic suppressing WFT oviposition. Certain acylsugar fatty acids, specifically i-C5, i-C9 and i-C11, were also significantly associated with changes in WFT oviposition. These results support continued breeding efforts to increase acylsugar amounts and explore modifications of fatty acid profile and their roles in deterring thrips oviposition. The finding that acylsugar production occurs and reduces thrips oviposition in tomato flowers will be important in efforts to use acylsugar-mediated resistance to reduce incidence of orthotospoviruses such as TSWV in tomato by deterring virus transmission and development of thrips vector populations in the crop.


Assuntos
Quimera/metabolismo , Ácidos Graxos/metabolismo , Flores , Melhoramento Vegetal , Solanum lycopersicum , Açúcares/metabolismo , Tisanópteros , Animais , Metabolismo dos Carboidratos/fisiologia , Cruzamentos Genéticos , Flores/metabolismo , Herbivoria , Insetos/efeitos dos fármacos , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Oviposição/genética , Controle Biológico de Vetores , Doenças das Plantas/genética , Doenças das Plantas/prevenção & controle , Plantas Geneticamente Modificadas , Açúcares/análise , Açúcares/farmacologia , Tisanópteros/genética , Tisanópteros/metabolismo
11.
Virology ; 493: 39-51, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-26999025

RESUMO

Tomato spotted wilt virus (TSWV) is transmitted by thrips in a propagative manner; however, progression of virus infection in the insect is not fully understood. The goal of this work was to study the morphology and infection of thrips salivary glands. The primary salivary glands (PSG) are complex, with three distinct regions that may have unique functions. Analysis of TSWV progression in thrips revealed the presence of viral proteins in the foregut, midgut, ligaments, tubular salivary glands (TSG), and efferent duct and filament structures connecting the TSG and PSG of first and second instar larvae. The primary site of virus infection shifted from the midgut and TSG in the larvae to the PSG in adults, suggesting that tissue tropism changes with insect development. TSG infection was detected in advance of PSG infection. These findings support the hypothesis that the TSG are involved in trafficking of TSWV to the PSG.


Assuntos
Tisanópteros/virologia , Tospovirus/fisiologia , Animais , Asteraceae/virologia , Datura stramonium/virologia , Feminino , Masculino , Glândulas Salivares/virologia , Tisanópteros/anatomia & histologia , Tospovirus/ultraestrutura
13.
PLoS One ; 9(4): e94447, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24736614

RESUMO

Saliva is known to play a crucial role in insect feeding behavior and virus transmission. Currently, little is known about the salivary glands and saliva of thrips, despite the fact that Frankliniella occidentalis (Pergande) (the western flower thrips) is a serious pest due to its destructive feeding, wide host range, and transmission of tospoviruses. As a first step towards characterizing thrips salivary gland functions, we sequenced the transcriptome of the primary salivary glands of F. occidentalis using short read sequencing (Illumina) technology. A de novo-assembled transcriptome revealed 31,392 high quality contigs with an average size of 605 bp. A total of 12,166 contigs had significant BLASTx or tBLASTx hits (E≤1.0E-6) to known proteins, whereas a high percentage (61.24%) of contigs had no apparent protein or nucleotide hits. Comparison of the F. occidentalis salivary gland transcriptome (sialotranscriptome) against a published F. occidentalis full body transcriptome assembled from Roche-454 reads revealed several contigs with putative annotations associated with salivary gland functions. KEGG pathway analysis of the sialotranscriptome revealed that the majority (18 out of the top 20 predicted KEGG pathways) of the salivary gland contig sequences match proteins involved in metabolism. We identified several genes likely to be involved in detoxification and inhibition of plant defense responses including aldehyde dehydrogenase, metalloprotease, glucose oxidase, glucose dehydrogenase, and regucalcin. We also identified several genes that may play a role in the extra-oral digestion of plant structural tissues including ß-glucosidase and pectin lyase; and the extra-oral digestion of sugars, including α-amylase, maltase, sucrase, and α-glucosidase. This is the first analysis of a sialotranscriptome for any Thysanopteran species and it provides a foundational tool to further our understanding of how thrips interact with their plant hosts and the viruses they transmit.


Assuntos
Perfilação da Expressão Gênica , Glândulas Salivares/metabolismo , Tisanópteros/genética , Ração Animal , Ontologia Genética , Genes de Insetos/genética , Anotação de Sequência Molecular , Análise de Sequência de DNA , Tisanópteros/virologia , Tospovirus/fisiologia
14.
Commun Integr Biol ; 5(1): 43-9, 2012 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-22482009

RESUMO

The majority of plant viruses rely on insect vectors for transmission. Insects with piercing-sucking mouthparts are the most common and efficient vectors because, they are able to inject viruses into specific plant tissues. Acquisition and inoculation of viruses occurs during specific vector feeding behaviors, and feeding behavior varies greatly among insects with piercing-sucking mouthparts. In this review we provide an overview of the feeding behavior of the major insect vectors with piercing sucking mouthparts: aphids, whiteflies, mealybugs, hoppers, and thrips. We briefly review the different mechanisms of plant virus transmission by these insects, and discuss how each mechanism requires a vector that engages in specific feeding behaviors, and how differences in feeding behavior among these insects can determine which viruses they are capable of transmitting. We also discuss recent findings indicating that plant viruses can directly modify their vector's behavior in a way that enhances transmission to a host plant.

15.
Proc Natl Acad Sci U S A ; 108(23): 9350-5, 2011 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-21606372

RESUMO

Vector infection by some animal-infecting parasites results in altered feeding that enhances transmission. Modification of vector behavior is of broad adaptive significance, as parasite fitness relies on passage to a new host, and vector feeding is nearly always essential for transmission. Although several plant viruses infect their insect vectors, we have shown that vector infection by a plant virus alters feeding behavior. Here we show that infection with Tomato spotted wilt virus (TSWV), type member of the only plant-infecting genus in the Bunyaviridae, alters the feeding behavior of its thrips vector, Frankliniella occidentalis (Pergande). Male thrips infected with TSWV fed more than uninfected males, with the frequency of all feeding behaviors increasing by up to threefold, thus increasing the probability of virus inoculation. Importantly, infected males made almost three times more noningestion probes (probes in which they salivate, but leave cells largely undamaged) compared with uninfected males. A functional cell is requisite for TSWV infection and cell-to-cell movement; thus, this behavior is most likely to establish virus infection. Some animal-infecting members of the Bunyaviridae (La Crosse virus and Rift Valley fever virus) also cause increased biting rates in infected vectors. Concomitantly, these data support the hypothesis that capacity to modify vector feeding behavior is a conserved trait among plant- and animal-infecting members of the Bunyaviridae that evolved as a mechanism to enhance virus transmission. Our results underscore the evolutionary importance of vector behavioral modification to diverse parasites with host ranges spanning both plant and animal kingdoms.


Assuntos
Comportamento Alimentar/fisiologia , Insetos Vetores/fisiologia , Insetos/fisiologia , Tospovirus/fisiologia , Animais , Bunyaviridae/fisiologia , Feminino , Interações Hospedeiro-Patógeno , Insetos Vetores/virologia , Insetos/virologia , Solanum lycopersicum/parasitologia , Solanum lycopersicum/virologia , Masculino , Folhas de Planta/parasitologia , Folhas de Planta/virologia , Fatores Sexuais , Fatores de Tempo
16.
Environ Entomol ; 38(4): 1069-77, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19689885

RESUMO

Bacterial populations in Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) collected in diverse California environments consisted of two bacterial symbionts: BFo-1 and BFo-2 (B = bacteria, Fo = Frankliniella occidentalis, numbers reflect different types). Dual infections of BFo-1 and BFo-2 were found in 50% of the thrips, 18% had neither bacterium, and 24 and 8% were infected solely with BFo-1 and BFo-2, respectively. No other bacteria consistently infected F. occidentalis. Dual infections occurred more often in male thrips and in thrips of both sexes from southern mountain and valley sites. As average collection year or month minimum temperature decreased, infections of BFo-1, alone or in dual infections, increased significantly. As yearly precipitation increased, infection with BFo-1 alone also increased. F. occidentalis color morphology did not affect bacterial infection. BFo-1 created weak biofilms at 25 and 32 degrees C; BFo-2 made strong biofilms at 25 degrees C and no biofilms at 32 degrees C. When the bacteria were grown in culture together, weak biofilms formed at both temperatures studied, although there was no way to determine what each bacterium contributed to the biofilm. BFo-1 and BFo-2 grew at similar rates at 25 and 30 degrees C. Our data show BFo-1 and BFo-2 occur in natural populations of F. occidentalis and support the hypothesis BFo have a symbiotic relationship with F. occidentalis. Regional differences in bacterial prevalence suggest bacterial infection is associated with environmental conditions, and altitude, temperature, and precipitation may be important factors.


Assuntos
Ecossistema , Insetos/microbiologia , Simbiose , Animais , Biofilmes , California , Fenótipo , Temperatura
17.
Phytopathology ; 99(4): 404-10, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19271982

RESUMO

Tomato spotted wilt virus (TSWV) is transmitted in a persistent propagative manner by Frankliniella occidentalis, the western flower thrips. While it is well established that vector competence depends on TSWV acquisition by young larvae and virus replication within the insect, the biological factors associated with frequency of transmission have not been well characterized. We hypothesized that the number of transmission events by a single adult thrips is determined, in part, by the amount of virus harbored (titer) by the insect. Transmission time-course experiments were conducted using a leaf disk assay to determine the efficiency and frequency of TSWV transmission following 2-day inoculation access periods (IAPs). Virus titer in individual adult thrips was determined by real-time quantitative reverse transcriptase-PCR (qRT-PCR) at the end of the experiments. On average, 59% of adults transmitted the virus during the first IAP (2 to 3 days post adult-eclosion). Male thrips were more efficient at transmitting TSWV multiple times compared with female thrips of the same cohort. However, females harbored two to three times more copies of TSWV-N RNA per insect, indicating that factors other than absolute virus titer in the insect contribute to a successful transmission event. Examination of virus titer in individual insects at the end of the third IAP (7 days post adult-eclosion) revealed significant and consistent positive associations between frequency of transmission and virus titer. Our data support the hypothesis that a viruliferous thrips is more likely to transmit multiple times if it harbors a high titer of virus. This quantitative relationship provides new insights into the biological parameters that may influence the spread of TSWV by thrips.


Assuntos
Insetos/virologia , Doenças das Plantas/virologia , RNA Viral/análise , Tospovirus/fisiologia , Animais , Datura/virologia , Feminino , Interações Hospedeiro-Patógeno , Masculino , Fatores Sexuais , Tospovirus/isolamento & purificação
18.
Mol Plant Microbe Interact ; 19(4): 383-8, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16610741

RESUMO

The Mi-1.2 gene in tomato (Solanum lycopersicum) is a member of the nucleotide-binding leucine-rich repeat (NBLRR) class of plant resistance genes, and confers resistance against root-knot nematodes (Meloidogyne spp.), the potato aphid (Macrosiphum euphorbiae), and the sweet potato whitefly (Bemisia tabaci). Mi-1.2 mediates a rapid local defensive response at the site of infection, although the signaling and defensive pathways required for resistance are largely unknown. In this study, eggplant (S. melongena) was transformed with Mi-1.2 to determine whether this gene can function in a genetic background other than tomato. Eggplants that carried Mi-1.2 displayed resistance to the root-knot nematode Meloidogyne javanica but were fully susceptible to the potato aphid, whereas a susceptible tomato line transformed with the same transgene was resistant to nematodes and aphids. This study shows that Mi-1.2 can confer nematode resistance in another Solanaceous species. It also indicates that the requirements for Mi-mediated aphid and nematode resistance differ. Potentially, aphid resistance requires additional genes that are not conserved between tomato and eggplant.


Assuntos
Genes de Plantas/genética , Nematoides/fisiologia , Doenças das Plantas/genética , Doenças das Plantas/parasitologia , Solanum lycopersicum/genética , Solanum melongena/genética , Solanum melongena/fisiologia , Animais , Afídeos/fisiologia , Interações Hospedeiro-Parasita , Filogenia , Plantas Geneticamente Modificadas
19.
Phytopathology ; 96(8): 819-27, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18943746

RESUMO

ABSTRACT RNA silencing has been shown to be an important mechanism for conferring resistance in transgenic, virus-resistant plants. We used this approach to evaluate resistance in Nicotiana benthamiana plants transformed with chimeric coding and noncoding sequences from Citrus tristeza virus (CTV). Several independent transgenic plant lines were generated, using two constructs (pCTV1 and pCTV2) designed to produce self-complementary transcripts. The pCTV1 contained cDNA sequences from the CTV capsid protein (CP), p20, and 3' untranslated region (UTR); and pCTV2 contained CP, p23, and 3' UTR sequences. Heterologous recombinant Potato virus X (PVX) containing either homologous or heterologous CTV sequences was used to challenge plants and resistance was evaluated phenotypically and validated with reverse-transcriptase polymerase chain reaction and northern hybridization analysis. Transgenic plants (T1 generation) for each construct showed resistance to recombinant PVX constructs used for challenge experiments when PVX contained p20 or UTR (for CTV1 plants), or p23 or UTR (for CTV2 plants). However, no resistance was seen when plants were challenged with PVX containing the CTV CP. T2 generation plants also showed resistance even when challenged with PVX containing the cognate CTV sequences obtained from heterologous CTV isolates. The presence of transgene-specific small interfering RNAs in the resistant CTV1 and CTV2 plants indicated that resistance was mediated by post-transcriptional gene silencing.

20.
Annu Rev Phytopathol ; 43: 459-89, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16078892

RESUMO

The complex and specific interplay between thrips, tospoviruses, and their shared plant hosts leads to outbreaks of crop disease epidemics of economic and social importance. The precise details of the processes underpinning the vector-virus-host interaction and their coordinated evolution increase our understanding of the general principles underlying pathogen transmission by insects, which in turn can be exploited to develop sustainable strategies for controlling the spread of the virus through plant populations. In this review, we focus primarily on recent progress toward understanding the biological processes and molecular interactions involved in the acquisition and transmission of Tospoviruses by their thrips vectors.


Assuntos
Insetos/fisiologia , Insetos/virologia , Doenças das Plantas/parasitologia , Doenças das Plantas/virologia , Tospovirus/fisiologia , Animais , Evolução Biológica , Interações Hospedeiro-Parasita , Insetos/genética , Tospovirus/genética
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